Separating the male
An experienced male will be the first one to indicate imminent Heat in one of "his" bitches because he constantly "tests" (smells and licks vulva) all his girls every day.
With a younger, inexperienced male, it will most likely be one of the older bitches who will indicate. (Usually by showing more interest, perhaps a little play-aggression, or extra jealousy when that bitch seeks attention. (bitches in Heat are often more affectionate and attention-seeking)). Soon after, all the girls and the male will check her out!
I will generally separate the male early on, as the inevitable pre-mounting behavior (male-female, female-male and/or female-female) is a huge aggression trigger within the pack dynamics.
After that, it is subjective when to separate the Heat Bitch but NEVER leave her without supervision in the Pack.
Separating the Heat Bitch
Regardless of a Bitch's standing in Pack heirarchy, Hormonal Changes Influence Behavioural Changes!
The Alpha bitch will assert her dominance more forcefully. (In Heat or not)
The 2ic bitch, if in Heat, will vie for Alpha position (since, potentially, she is about to become (pregnant) Queen.
For any of the non-dominant bitches, Heat can bring on aggressive/temperament changes not normally apparent.
For All Pack bitches, this is a time of unsettled behavior. They may all align with the Dominant who suddenly tries to oust the 2ic from the Pack.
Play can take on a more aggressive edge.
Feed times can erupt into "bowl-hopping".
Mounting behaviours can turn aggressive.
(Mounting is a natural part of both Dominance-play and Mating-ritual learning)
Rule Of Thumb
At the first sign of aggression, separate.. there will be no 2nd chances.
If 2 bitches fight once, they will ALWAYS fight!
So, how long to keep separated?
For the male, most likely a week post-ovulation (wash the bitch and clean the house. This won't remove the scent that dogs can still smell but it will dampen it enough not to trigger the male's territory-marking)
For bitches, this will depend on normal Pack dynamics, but potentially 3 to 5 days post-ovulation.
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